Methods and compositions for personalized medicine by point-of-care devices for brain natriuretic peptide

ABSTRACT

Methods, devices, reagent, systems and kits for the detection, diagnosis of ovarian cancer as well as for the monitoring of ovarian cancer progression and for monitoring the progress of ovarian cancer treatments using BNP as a biomarker.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a division of U.S. application Ser. No. 14/526,376,filed

Oct. 28, 2014, which claims the benefit of U.S. Provisional ApplicationNo. 61/961,969, filed Oct. 28, 2013, and is a continuation-in-part ofPCT/US2013/046040, filed Jun. 15, 2013, which claims the benefit of U.S.Provisional Application No. 61/659,981, filed Jun. 15, 2012; U.S.Provisional Application No. 61/667,081, filed Jul. 2, 2012; and U.S.Provisional Application No. 61/671,717, filed Jul. 14, 2012, eachexpressly incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to methods, devices, reagent, systems andkits for the detection, diagnosis of ovarian cancer as well as for themonitoring of ovarian cancer progression and for monitoring the progressof various cancer treatments including ovarian cancer. The presentinvention also relates to point-of-care testing (POCT) and methods fordetermining concentrations of brain natriuretic peptide (BNP) in asubject.

BACKGROUND OF THE INVENTION

Cancer is a leading cause of death worldwide, accounting for 7.6 milliondeaths (around 13% of all deaths) in 2008. Ovarian cancer is the ninthmost common cancer in women and the fifth leading cause ofcancer-related deaths in women in the US. One of every 72 women willdevelop ovarian cancer and one of every 100 will die from this form ofcancer. The American Cancer Society estimates that in 2013, 22,240 womenwill be diagnosed with ovarian cancer and about 14,230 will die fromovarian cancer. About 85% to 90% of ovarian cancers are epithelialovarian carcinomas.

Treatment options include surgery, chemotherapy, and occasionallyradiation therapy. Surgery usually involves the removal of one or bothovaries, fallopian tubes and the uterus. In advanced disease, surgicallyremoving all abdominal metastases enhances the effect of chemotherapyand helps improve survival. For women with stage III ovarian cancer inwhich removal of cancerous tissue has been performed, studies show thatchemotherapy administered both intravenously and directly into theperitoneal cavity improves survival.

The identification of tumor markers suitable for early detection anddiagnosis of cancer and in particular, ovarian cancer would improve theclinical outcome of patients, especially those presenting vague or nosymptoms. Presently no cost effective screening tests have beendeveloped.

Ovarian epithelial cancer is more common in individuals with elevatedgonadotropin-releasing hormone (GnRH) including follicle-stimulatinghormone (FSH) and luteinizing hormone (LH), such as postmenopausal womenor women who have received treatment to induce ovulation. Conversely,reduced risk of ovarian cancer is associated with a history of multiplepregnancies, breastfeeding, oral contraceptive use, and estrogenreplacement therapy, all of which are related to lower levels of andreduced exposure to FSH and LH. FSH, follicle stimulating hormone,regulates gene expression in ovarian tumors (Chu S, Rushdi S, Zumpe ET,Mamers P, Healy DL, Jobling T, Burger HG, Fuller PJ. (2002)FSH-regulated gene expression profiles in ovarian tumors and normalovaries. Mol Hum Reprod. 8:426-33) and causes neovascularization ofovarian cancers by increasing vascular endothelial growth factor (VEGF)expression through upregulation of surviving (Huang Y, Hua K, Zhou X,Jin H, Chen X, Lu X, Yu Y, Zha X, Feng Y. (2008) Activation of thePI3K/AKT pathway mediates FSH-stimulated VEGF expression in ovarianserous cystadenocarcinoma. Cell Res. 18:780-91).

Currently, cancer antigen 125 (CA-125) is used as a serum biomarker forovarian cancer. Serum concentrations of CA-125 are elevated in 75-80% ofpatients with advanced-stage disease and this marker. CA125 is used as aserum tumor marker for monitoring response to chemotherapy, detectingdisease recurrence, as well as distinguishing malignant from benignpelvic masses. However, it is presently not an appropriate diagnosticbiomarker as the majority of healthy women with high levels of CA-125 donot have cancer.

Accordingly, there is a need for improved methods of detection anddiagnosis of cancer including ovarian cancer as well as methods formonitoring the progress of the disease and monitoring the progress ofvarious treatments for ovarian cancer including point of care or pointof use devices capable of quantitating predictive biomarker(s).

SUMMARY OF THE INVENTION

The present invention relates methods, devices, reagent, systems andkits for the for the detection and diagnosis of ovarian cancer usingbrain natriuretic peptide (BNP) as a biomarker.

The present invention relates to methods, devices, reagent, systems andkits for monitoring the progression of ovarian cancer and for monitoringthe progress of ovarian cancer treatments.

The present invention also relates to quantitative point-of-care devicesand methods for the detection of BNP as a diagnostic for ovarian cancer.

In certain embodiments, the present invention provides a method formonitoring BNP for an individual treated with a drug. The methodinvolves obtaining samples from the individual at suitable time points.The samples may be collected at point-of-care or point-of-use bysampling or self-sampling on point-of-care devices or point of usedevices, each capable of quantitating the biomarker, or on matricessuitable for storage of at least two samples prior to quantitation ofthe drug by a central laboratory. The information obtained may besuitable for guiding dosing of the drug for the individual.

The invention also relates to point-of-care and/or point-of-use devicesfor quantitation BNP which allows for personalized dosing and monitoringfor more effective therapies for ovarian cancer.

Samples may be collected by at point-of-care or point of service, e.g.,by self-sampling. Samples may be applied to a lateral flow device forquantitation of the drug, and the results transmitted to the physicianor physician's agent for pharmacokinetic analysis. In other embodiments,the samples are collected at point-of-care or point-of-service, e.g., byself-sampling, on a suitable storage matrix, e.g., nitrocellulose, priorto delivery of the samples to a central laboratory for quantitation andanalysis.

In certain embodiments, samples collected at various times from theindividual through point-of-care or point-of-use by self-sampling may beobtained by a central laboratory. The laboratory then tests the samplesto quantitate the biomarker of interest and, based on the results,detection or diagnosis of ovarian cancers may be obtained. The resultsobtained may also be used to determine the magnitude of the diseases'progression as well as to monitor the efficacy of treatment regimens.

In another aspect it provides a kit for biomarker monitoring of anindividual treated with a drug. The kit comprises a plurality ofpoint-of-care device or a point of use device capable of quantitatingthe drug in one or more samples, or matrices suitable for storage of thesamples prior to quantitation by a central laboratory.

DESCRIPTION OF THE DRAWINGS

The foregoing aspects and many of the attendant advantages of thisinvention will become more readily appreciated as the same become betterunderstood by reference to the following detailed description, whentaken in conjunction with the accompanying drawings.

FIGS. 1A-1C are plots of BNP concentration versus CA125 concentration(1A), FSH concentration versus CA125 concentration (1B), and BNPconcentration versus FSH concentration (1C).

FIGS. 2A-2D are plots of age matched (2A) (donor 50 yr; ovarian cancerpatient 51 yr), height matched (2B) (donor 166 cm; ovarian cancerpatient 165 cm), body mass index (BMI) (2C) (donor 25; ovarian cancerpatient 27), and weight matched (2D) (donor 70 kg; ovarian cancerpatient 72 kg) donors and ovarian cancer patients.

FIG. 3 is a plot of the BNP levels (ng/mL) of ovarian cancer patientsand donors (t Test Prob>F<0.0001) (donor 126.35 ng/mL; ovarian cancerpatient 55.34 ng/mL).

FIG. 4A is a bar graph of number of patients (79) and donors (55) withhypertension (patients with hypertension 30; donors with hypertension 1)and without hypertension (patients without hypertension 49; donorswithout hypertension 54).

FIG. 4B is a plot of BNP levels in ovarian cancer patients withhypertension (Y) (30 patients; mean 68.9423) and without hypertension(N) (49 patients; mean 64.0855) (t Test Prob>F<0.7269).

FIG. 5 is a plot of BNP levels in ovarian cancer patients (49 patients;mean 68.881) and donors (44 donors; mean 171.066) who are normotensive(t Test Prob>F<0.0001).

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to methods, devices, reagent, systems andkits for the detection, diagnosis, and progression of ovarian cancerusing brain natriuretic peptide (BNP) as a biomarker.

Biomarker data may be obtained from samples collected at point-of-careor point-of-use using for example a lateral flow point of care test.Advantageously, the samples may be obtained by self-sampling. In certainembodiments, the samples may be delivered to a point-of-care device toquantitate the biomarker, and the results thus obtained are reported tothe physician or his agent. In one embodiment quantitation of brainnatriuretic peptide (BNP) can be monitored using point-of-care andpoint-of-use detection which allows for personalized dosing andmonitoring for more effective therapies for various cancers includingovarian cancers. BNP may be quantitatively detected from various bodilyfluids including but not limited to plasma, serum or urine.Point-of-care needs to have an expanded dynamic range of concentrationdetection. Preferably the assays or test require a single determinationwith no repeat and no dilution at point-of-care. Most immunoassays havea working range of 2 logs. The methods of the present invention providefor expanding the range of detection for the biomarkers at least one logor at least two logs or at least 3 logs or at least 4 logs or at least 5logs or at least 6 logs or at least 7 logs or at least 8 logs. Forexample the assays and tests and methods of the present invention permitan expanded dynamic range of detectable concentrations of BNP. Theexpanded dynamic range encompasses all possible concentrations of BNPencountered in blood, serum or other bodily fluids. The assays and testsand methods of the present invention also provide cassettes which can beread using an optical reader with 2D barcode capability, and providethat the data can be printed out or stored on the reader for uploadingonto a database including but not limited to a clinic, doctor's officeor hospital database. The reader utilizes confocal optics with a lowdistance-to-target ratio. The reflectometric measurement is converted toactivity units, using an established calibration curve embedded in the2D barcode. The cassettes are made such that the samples within arestable for at least 12 or at least 24 or at least 48 or at least 72hours and thus can be shipped to a central lab or doctor's office forquantitation if the patient does not have access to the reader.

The readers utilized in the present invention include but are notlimited to confocal optical readers or cell/smart phone readers. Areflectometric optical reader, which utilizes confocal optics with a lowdistance-to-target ratio, may be used with the methods, kits and assaysof the present inventions. Calculations may be performed in thebackground using information embedded on the 2D-bar code specific toeach lot of cassettes. Alternatively a cell phone reader which providesquantitation at a point-of-care without sufficient resources can be usedto capture images of the cassettes and transmit the images over theinternet to a facility such as a centralized processing facility wherethe BNP values can be received in real time.

The present invention provides rapid and quantitative point-of-caretesting for BNP including field deployment directly at home or inemergency care situations, and provides for cassettes which can be readdirectly by the patient or can be shipped to the central lab/doctor'soffice for reading. As therapeutic drug monitoring (TDM), the testshould allow for more effective dosing of the patients and therebyimproving effectiveness of treatment. The tests are alsopatient-centric, inviting better compliance and patient participation inpersonalizing his/her treatment. The simplicity of the assays wouldallow for their deployment in underdeveloped regions lacking access tocentral laboratories with specialized and expensive equipment. Theexpanded range eliminates the need for dilution of the samples to bringthem within working range of the traditional assay.

Alternatively, the samples are collected using a matrix or vesselsuitable for collection and storage of the samples until receipt andanalysis by a central laboratory. Examples of matrices or vesselssuitable for collection and storage of the samples include, but are notlimited to, commercially available biological sampling filter papersystems, such as WHATMAN® 3 MM, GF/CM30, GF/QA30, S&S 903, GB002, GB003,or GB004. Several categories of blotting materials for blood specimencollection are available, e.g., S&S 903 cellulose (wood or cottonderived) filter paper and WHATMAN® glass fiber filter paper. The bloodspot is placed in one or more designated areas of the filter paper,allowed to dry, and then mailed along with a test request form to thecentral laboratory. This method of collection has the advantage ofobviating the need for collection of samples at a doctor's office orclinic. Thus, multiple samples may be conveniently collected by thepatient over a period of 0 to 72 hours at considerable savings of costand time. This has the advantages of increased efficiency and reduceddelays in transmitting results of the analysis to the treatingphysician, who may use the information to adjust treatment as necessary,and contact the patient to convey the new treatment regimen. In oneaspect, one or more biomarkers are provided for use either alone or invarious combinations to diagnose ovarian cancer, permit differentialdiagnosis of pelvic masses as benign or malignant, monitor ovariancancer progression or monitor ovarian cancer recurrence.

Any of the biomarkers described herein may be used in a variety ofclinical indications for ovarian cancer, including any of the following:detection of ovarian cancer, characterizing ovarian cancer (e.g.,determining ovarian cancer type, sub-type or stage), such as bydetermining whether a pelvic mass is benign or malignant; determiningovarian cancer prognosis; monitoring ovarian cancer progression orremission; monitoring for ovarian cancer recurrence; monitoringmetastasis; treatment selection (e.g., pre- or post-operativechemotherapy selection; monitoring response to a therapeutic agent orother treatment, combining biomarker testing with additional biomedicalinformation).

As an example of the manner in which the biomarker described herein maybe used to diagnose ovarian cancer, differential expression of one ormore of the biomarkers described herein in an individual who is notknown to have ovarian cancer may indicate that the individual hasovarian cancer thereby enabling detection of ovarian cancer at an earlystage of the disease when treatment is most effective. Decreasedexpression of the biomarker from “normal” during the course of ovariancancer may be indicative of ovarian cancer progression whereas anincrease in the expression as compared with normal expression mayindicate that the individual is in remission or is being successfullytreated. Decreases in the degree of biomarker expression as compared to“normal” may indicate cancer progression or ineffectiveness of ovariancancer treatment. Additionally a decrease in the differential expressionof one or more of the biomarkers after an individual has apparently beencured of ovarian cancer may be indicative or ovarian cancer recurrence.In this case, cancer treatment may be resumed or current treatment maybe augmented or supplemented as need be. In addition, a differentialchange in the level of biomarker might also be indicative of anindividual's response to a particular therapeutic agent. Differentialexpression refers to expression of a biomarker that is activated to ahigher or lower level in a subject suffering from a specific disease,relative to its expression in a normal or control subject or a subjectwho does not have the specific disease. Differential expression includesboth quantitative and qualitative differences in expression among normaland diseased cells or among cells which have undergone different diseaseevents or different treatments.

The biomarker of the present invention is brain natriuretic peptide(BNP). Brain natriuretic peptide is a 32 aminoacid, approximately 3 kDapeptide that is encoded by the human NPPB gene and plays a role in themodulation of diuresis, vasorelaxation, and secretion of renin andaldosterone.

The BNP biomarker may be used in combination with one or more otherbiomarkers for ovarian cancer or other diseases or conditions (e.g.,other cancers). Thus, the BNP biomarker can be used as a biomarker in abiomarker panel.

The BNP biomarker may be differentially expressed at any level, but isgenerally present at a level that is decreased by at least 5%, by atleast 10%, by at least 15%, by at least 20%, by at least 25%, by atleast 30%, by at least 35%, by at least 40%, by at least 45%, by atleast 50%, by at least 55%, by at least 60%, by at least 65%, by atleast 70%, by at least 75%, by at least 80%, by at least 85%, by atleast 90%, by at least 95%, or by 100%. The level of expression of thebiomarker of the present invention may be between about 10% lower toabout 100% lower or about 10% lower to about 90% lower or about 10%lower to about 80% lower or about 10% lower to about 70% lower or fromabout 10% lower to about 60% lower or from about 10% lower to about 50%lower or from about 10% lower to about 40% lower or from about 10% lowerto about 30% lower or about 10% lower to about 20% lower or about 20%lower to about 100% lower or about 20% lower to about 90% lower or fromabout 20% lower to about 80% lower or from about 20% lower to about 70%lower or from about 20% lower to about 60% lower or from about 20% lowerto about 50% lower or from about 20% lower to about 40% lower or fromabout 20% lower to about 30% lower or from about 50% lower to about 100%lower than normal or than that of a control.

The biomarker is preferably differentially present at a level that isstatistically significant (e.g., a p-value less that 0.05 and/or aq-value of less than 0.10 as determined by either Welch's T-test orWilcoxon's rank-sum test). Alternatively, the biomarker demonstrates acorrelation with the presence of ovarian cancer or particular stages ofovarian cancer. The range of correlations is between negative 1 (−1), aperfect negative correlation, and positive 1 (+1), a perfect positivecorrelation. Zero (0) would mean no correlation. A substantial positivecorrelation refers to a biomarker having a correlation between +0.25 and+1.0 with a disease or clinical measurement while a substantial negativecorrelation refers to a biomarker having a correlation between −0.25 and−1.0 with a disease or clinical measurement. A significant positivecorrelation refers to a biomarker having a correlation between +0.25 and+1.0 with a disease or clinical measurement while a significant negativecorrelation refers to a biomarker having a correlation between −0.25 and−1.0 with a disease or clinical measurement.

In some cases it will be desirable to establish normal or baselinevalues (or ranges) for biomarker expression levels. Normal levels can bedetermined for any particular population, subpopulation or groupaccording to standard methods known to those of skill in the art.Generally baseline (normal) levels of biomarkers are determined byquantifying the amount of biomarker in biological samples (e.g., fluids,cells or tissues) obtained from normal (healthy) individuals.Application of standard statistical methods permits determination ofbaseline levels of expression as well as deviations from such baselinelevels.

A biomarker value can be detected by using any of a variety of knownanalytical methods. Biomarker detection may be facilitated by the use ofa capture agent which is one or more molecules which can specificallybind the biomarker. The capture agent, in solution or immobilized on asolid support, may be exposed to the biomarker and binding may bedetected in a variety of ways including, but not limited to,fluorescence, chemiluminescence, dyes, and other optically detectablemeans. Immunoassay methods are based on the binding of an antibody toits corresponding analyte (e.g., BNP) and can detect the analyte in asample depending on the specific assay format.

According to the American Cancer Society ovarian cancer can be stagedaccording to the AJCC/TNM System. This describes the extent of theprimary tumor (T), the absence or presence of metastasis to nearby lymphnodes (N), and the absence or presence of distant metastasis (M). Tcategories for ovarian cancer include: T1: the cancer is confined to oneor both ovaries; T2: the cancer is in one or both ovaries and isextending into pelvic tissues; and T3: the cancer is in one or bothovaries and has spread to peritoneum. N categories indicate if thecancer has spread to regional (nearby) lymph nodes. Nx: no descriptionof lymph node involvement is possible because information is incomplete.NO: no lymph node involvement. N1: cancer cells are found in the lymphnodes close to tumor. Once a patient's T, N, and M categories have beendetermined, this information is combined in a process called stagegrouping to determine the stage, expressed in Roman numerals from stageI (the least advanced stage) to stage IV (the most advanced stage).Stage 1: the cancer is still contained within the ovary (or ovaries). Ithas not spread outside the ovary. Stage 2: the cancer is in one or bothovaries and has spread to other organs (such as the uterus, fallopiantubes, bladder, the sigmoid colon, or the rectum) within the pelvis. Ithas not spread to lymph nodes, the peritoneum, or distant sites. Stage3: the cancer is in one or both ovaries, and one or both of thefollowing are present: (1) cancer has spread beyond the pelvis to thelining of the abdomen; (2) cancer has spread to lymph nodes. Stage 4:the most advanced stage of ovarian cancer; in this stage the cancer hasspread to the inside of the liver, the lungs, or other organs locatedoutside the peritoneal cavity. Finding ovarian cancer cells in the fluidaround the lungs is also evidence of stage IV disease.

Non-limiting examples of suitable devices or methods of testing drugsinclude lateral flow devices for the determination of the concentrationof an analyte in a sample comprising providing a lateral flow strip foruse in measuring the analyte. Examples of analytes that may be testedinclude therapeutic drugs, drug metabolites, and hormones. Applicationof the sample to the lateral flow strip causes a fraction of the analytein the sample to bind to a component of the lateral flow strip such thata detectable signal proportional to the concentration of the analyte inthe sample is produced.

Alternatively, the quantitation may be conducted on samples submitted byindividuals to a laboratory by any suitable assay, including, but notlimited to, those currently known to the art, such as ELISA, liquidchromatography-mass spectrometry (LC-MS), thin layer chromatography(TLC), high-performance liquid chromatography (HPLC), and massspectrometry (MS) or other traditional assays for drug monitoring atcentral lab have been well illustrated. The samples could be whole bloodcollected following a finger prick on a suitable matrix and stored as adry blood spot that is shipped or otherwise delivered to a laboratoryfor testing. Sampling can be performed with capillary and/or devicedesigned to deliver precise and small amount of blood to the dried bloodspot card- card punch variability replaced pipette variability.

The following examples are provide for the purpose of illustrating, notlimiting the invention.

EXAMPLE 1 Measurement of LH, FSH, and BNP Levels

Serum samples collected at time of diagnosis of ovarian cancer weretested using rapid and quantitative point-of-care (POC) devices forblood biomarkers (LH, FSH, and BNP) and the data was evaluated usingJMP9 statistical analysis software. Quantitative lateral flow assays forFSH and LH were performed according to Larn Hwang, Chao Hsiao, KourosMotamed, Vuong Trieu (2012) Rapid and Quantitative Lateral FlowPoint-of-Care Therapeutic Drug Monitoring (TDM) Assays for LH and FSH.American Association for Cancer Research (AACR) Annual Meeting, Mar.31-Apr. 4, 2012. Quantitative BNP assay was from Humasis (Korea). FSHrange=5-10,000 IU/L; LH range=1-1,700 IU/L; BNP range=25-800pg/mL.

TABLE 1 BNP (pg/mL) Quartiles 10% 25% median 75% 95% Normal 0 0 0 84.1177.3 Ovarian Cancer 0 0 40.0 74.8 127.3 p = 0.02, chi-square

TABLE 2 LH (pg/mL) Quartiles 10% 25% median 75% 95% Normal 4.83 6.0016.65 62.53 83.72 Ovarian Cancer 4.50 12.60 17.40 40.00 61.20 p = 0.9817Wilcoxon statistics

TABLE 3 BNP (pg/mL) Quartiles 10% 25% median 75% 95% Normal 3.79 7.7513.40 99.68 828.84 Ovarian Cancer 15.10 74.30 151.60 418.60 825.00 p =0.0102 Wilcoxon statistics

In the serous adenocarcinoma group, FSH level was higher (median=151.6mU/ml) vs. normal controls (median of 13.4 mU/ml, p=0.01, Wilcoxon).Moreover, incidence of BNP >25 pg/ml was higher for patients (14 of 19,74%) vs. normal controls (3 of 10, 30%, p=0.02, Chi-square). FSHprogressively increased from normal controls, to normotensive patients,to hypertensive patients with median FSH values of 13.4, 79.3, and232.2, respectively. The same was not observed for BNP. FSH increase inhypertensive patients was not accompanied by increase in BNP. However,incidence of BNP>25 pg/ml was higher for patients (14 of 19, 74%) vs.normal controls (3 of 10, 30%, p=0.02, Chi-square). No differences wereobserved for LH. These data suggest the BNP and FSH hormones play arole(s) in ovarian cancer.

There were no correlations between CA125 and FSH or BNP suggesting thatthey are independent biomarkers (see FIGS. 1A-1C). FSH and BNP exhibitedincreased incidence/level among cancer patients versus age matchednormal. FSH is probably acting as regulator of cancer cell expression aswell as induction of angiogenesis. Elevated levels of FSH are shown inFIG. 1B. The role of BNP is unknown at the moment. The data shown inthis example demonstrated the use of POC/POU device for monitoring therelevant blood biomarkers (FSH, LH, hCG, and BNP). The association ofBNP with ovarian cancer was previously unrecognized.

EXAMPLE 2 Measurement of BNP levels in Ovarian Cancer Patients

BNP levels were determined for 55 donors and 79 ovarian cancer patients.The donors and ovarian cancer patients were matched for age (donor 50years; patient 51 years), height (donor 166 cm; patient 165 cm), weight(donor 70 kg; patient 72 kg) and BMI (donor 25; patient 27) as seen inFIGS. 2A-2D. The donor average BNP was 126.36 ng/mL, whereas the averageBNP level for the patients was 55.34 ng/mL. FIG. 3 shows a plot of thedata.

As the ovarian cancer population was imbalanced for hypertension, asubgroup analysis was performed. Of the 55 donors, one had hypertensionand in the patient group 30 had hypertension and 49 were normotensive(FIG. 4A). The hypertensive and normotensive patients were compared(FIG. 4B) and the mean BNP level for the hypertensive patients was68.9423 ng/mL and the mean BNP level for the normotensive patients was64.0855 ng/mL. The normotensive patients were compared to normotensivedonors (FIG. 5). The mean BNP level for the normotensive patients was68.881 ng/mL and the mean BNP level for the normotensive donors was171.066 ng/mL. Thus, there is lower BNP in normotensive cancer patientsversus normotensive donors.

BNP level is depressed in ovarian cancer patients and there appears tobe little to no differences between hypertensive patients versusnormotensive patients with respect to BNP levels of those with ovariancancer. BNP level remained depressed when normotensive patients werecompared to normotensive donors. This is the first demonstration of alinkage between ovarian cancer and the cardiovascular system.

Where ranges are given herein, the endpoints are included. Furthermore,it is to be understood that unless otherwise indicated or otherwiseevident from the context and understanding of one of ordinary skill inthe art, values that are expressed as ranges can assume any specificvalue or subrange within the stated ranges in different embodiments ofthe invention, to the tenth of the unit of the lower limit of the range,unless the context clearly dictates otherwise.

All publications and patents cited in this specification are hereinincorporated by reference as if each individual publication or patentwere specifically and individually indicated to be incorporated byreference. The citation of any publication is for its disclosure priorto the filing date and should not be construed as an admission that thepresent invention is not entitled to antedate such publication by virtueof prior invention.

While this invention has been particularly shown and described withreferences to example embodiments thereof, it will be understood bythose skilled in the art that the various changes in form and detailsmay be made therein without departing from the scope of the inventionencompassed by the appended claims.

Further advantages of the methods of the present invention can beachieved by those skilled in the art based upon the embodimentsdescribed herein and are thus specifically within the scope of thepresent invention.

While illustrative embodiments have been illustrated and described, itwill be appreciated that various changes can be made therein withoutdeparting from the spirit and scope of the invention.

The embodiments of the invention in which an exclusive property orprivilege is claimed are defined as follows:
 1. A method for diagnosingthe presence of ovarian cancer in a subject, comprising determining thelevel of brain natriuretic peptide (BNP) in the subject and comparingthe level(s) of the biomarker(s) in the subject with the normal level ofBNP.
 2. The method of claim 1, wherein the level of biomarker in thesubject is at least 10% less than the normal level.
 3. The method ofclaim 1, wherein the level of biomarker in the subject is at least 50%less than the normal level.
 4. The method of claim 1, wherein thedetermination of the level of biomarker in the patient is performed by aquantitative point-of-care device.
 5. The method of claim 4, wherein thepoint-of-care device is a lateral flow device.
 6. A method formonitoring the progression of ovarian cancer in a patient diagnosed withovarian cancer, comprising determining the level of BNP in the patientat a first point in time and comparing the level of BNP in the subjectat a second point in time to determine the progression of the ovariancancer.
 7. The method of claim 6, wherein the level of biomarker in thesubject is at least 10% less than the normal level.
 8. The method ofclaim 6, wherein the level of biomarker in the subject is at least 50%less than the normal level.
 9. The method of claim 6, wherein thedetermination of the level of biomarker in the patient is performed by aquantitative point-of-care device.
 10. The method of claim 9, whereinthe POC device is a lateral flow device.
 11. A kit for biomarkermonitoring of an individual treated with an ovarian cancer treatment,the kit comprising: a plurality of point-of-care devices or apoint-of-use device capable of quantitating BNP in one or more bloodsamples.
 12. The kit of claim 11, further comprising instructions forcollecting the at least two samples.
 13. The kit of claim 11, whereinthe point-of-care device is an expanded range, lateral flow device.